EA022911B1 - Method for treatment of a patient suffering from i or ii stage of systemic lupus erythematosus - Google Patents

Abstract

In various embodiments, the present invention relates to a method for the treatment of a patient suffering from I or II stage of systemic lupus erythematosus (SLE), comprising weekly administering in a dosage from about 1 to about 10 mg/kg during about 2 to about 52 weeks of a fusion molecule which comprises TACI extracellular domain or fragment thereof which binds BiyS, and a human immunoglobulin-constant domain.

Description

In various embodiments, the present invention relates to methods and compositions for treating autoimmune diseases or disorders of the immune system, namely systemic lupus erythematosus, comprising administering a TASH-Gd fusion protein such as atacicept using a special dosage regimen that maximizes blocking the functions of the ΤΝΒ family of ligands .

BACKGROUND OF THE INVENTION

Ligand B1u8 / Family of receptors.

Three receptors, TAC1 (a transmembrane activator or transmembrane activator and partner of a calcium modulator and cyclophilin ligand), BCMA (B cell maturation antigen), and BAPP-K (receptor for B cell activating factor belonging to the ΤΝΡ family) were identified as those that have unique binding affinities for the two growth factors B1y8 (a stimulator of B-lymphocytes) and ARK1b (a ligand that induces proliferation) (Mag8! eg8 e! a1. Sigg Wu1 2000; 10 (13): 785-788; Tyotr8op e! A1.8c1epse 2001; 293: 2108-2111). TAC1 and BCMA are associated with both B1u8 and ARK1B. while BAPP-K was defined as one that associates with B1u8 with high affinity (Mag81eg8 e! a1. Sigg Vu1 2000; 10 (13): 785-788; Tyotrzop e! a1. 8c1epse 2001; 293: 21 08 -2111). As a result of this, B1y8 is capable of transmitting a signal through three receptors, while ARKGb is able to transmit a signal only through TAC1 and BCMA. In addition, the circulating heterotrimeric complexes B1y8 and ARKG (groups consisting of three proteins containing one or two copies of each of B1y8 and ARKG) were identified in serum samples taken from patients with systemic immune rheumatic diseases and were demonstrated as those that induce proliferation of клетокη ugo B cells (Co8syke e! a1. 1. Gttypo1 2002; 169: 4314-4321). Among the Gd-fusion proteins for all three receptors, only TASH-Pc5, such as ataccept, is capable of blocking the biological activity of heterotrimeric complexes (Cosier e! A1. 1. Gtipo1 2002; 169: 4314-4321).

B1u8 and ARKG are powerful stimulators of the maturation, proliferation and survival of B cells (Ogo88 e! A1. No. Zige 2000; 404: 995-999, Ogo88 e! A1. Gtttspyu 2001; 15 (2): 289-302, Ogoot e ! a1. 1. С1ш Гпуе8! 2002; 109 (1): 59-68). B1y8 and ARKHB may be necessary for the persistence of autoimmune diseases, in particular those that involve B cells in their development. Transgenic mice designed to express high levels of B1y8 exhibit disorders of the cells of the immune system, as well as symptoms similar to those observed in patients with systemic erythematic lupus erythematosus (Sje8op e! A1. Keu18ey dsheype8! Og1lpo818 apy 1gea! Tep! B1oy 1996; 87: 4990-4997, Sjeta e! A1. ΛΠΐιπΙί8 Kjeit 2001; 44 (6): 1313-1319). Similarly, elevated levels of B1y8 and ARKH were measured in serum samples taken from 8BE patients and other patients with various autoimmune diseases like rheumatoid arthritis (Co8ke e! A1. 1. Gtipo1 2002; 169: 4314-4321; Mape !! e X., App Kjeit Όΐ8 2003; 62 (2): 168-171; Naipe e! A1. 1. Exp May 1998; 188 (6): 11851190), which expanded the association of B1u8 and / or ARB with diseases that are mediated by Cells , from animal models to humans.

Systemic erythematous lupus erythematosus (8E) is an autoimmune disease that is clinically characterized by an increasing and dying out course and the involvement of numerous organs, including the skin, kidneys and central nervous system (Katteg OM and T8oko8 O.S. ed. (1999), Ltsrts8 : Mo1esi1ag apy Ce11i1ag Ra1yodepe818 1st ed., Nitap Рge88, Ν.Ι; Laiya K.O.d. (1999), 8u8! Etuuuuuuuuu! O8i8, 3rd ed., Asayetk Rge88, At8! yyyat). The total distribution of SbE is 1 in 2000, and approximately 1 out of 700 white women develop SbE during their life-time period (Laya K.O. (1999), Sigg. Orth. Kieita! O1. 8p; 11 (5): 352-6 ) In the United States alone, more than half a million people have 8LF, and most of them are women of reproductive age (NagLA. (2003), 1. Exp. May. 185: 1101-1111).

There is no single criterion for the diagnosis of 8E The American College of Rheumatology has developed 11 criteria for the diagnosis of 8E, which cover the clinical spectrum of 8E for skin, system and laboratory tests. Such criteria include rashes on the cheeks, discoid rashes, sensitivity to sunlight, ulcers in the oral cavity, arthritis, inflammation of the serous membranes, inflammation of the kidneys and central nervous system, changes in the composition of the blood, and the presence of antinuclear antibodies. The patient must meet four of these criteria in order to be diagnosed as an 8E patient. (Tap e! A1. (1982), AygJ18 Kieita! O1. 25: 1271-1277). 8bE is usually confirmed by assays that include, but are not limited to, blood tests to determine antinuclear antibodies; blood and urine tests to evaluate kidney function; analyzes using complement to determine the presence of low levels of complement, which is often associated with 8E; sedimentation rate (E8K) or the presence of C-reactive protein for measuring inflammation levels (SCR); radiography to assess lung damage and an ECG to assess heart damage.

Standard therapy for 8E is the administration of a steroidal glucocorticoid, a general inhibitor of the immune system. It can be used to relieve symptoms; however, no treatment for 8E is currently available. Low oral dosage is usually prescribed.

- 1,022,911 doses of prednisone at levels lower than 0.5 mg / kg / day. Unfortunately, this therapy is not enough to keep patients in a state of remission, and diseases often occur. Exacerbations can be controlled using high doses of glucocorticoid by intravenous administration at a dose of 30 mg methylprednisolone / kg / day for three consecutive days. However, steroid treatment at high doses can have severe side effects for patients.

Such standard treatments are generally non-specific, often associated with severe side effects and do not significantly affect the development of the disease or cause a transition to life-threatening kidney function complications (lupus nephritis or LF). Therefore, there is a long-felt need for the prior art in the development of new methods for treating 8E.

Summary of the invention

The present invention relates to a method for treating a patient suffering from stage I or II of systemic lupus erythematosus, which includes weekly administration at a dose of from about 1 to about 10 mg / kg for about 2 to about 52 weeks of a fusion molecule containing TAC1 extracellular domain or its a fragment that binds to B1y§ and the constant domain of human immunoglobulin.

Brief Description of the Figures

FIG. 1 graphically represents the concentration of free atacicept, plotted against the time, expressed in days, for subcutaneous administration, the main pharmacokinetic measurement. Each line in the graph represents a dose, as presented in the explanations.

FIG. 2 graphically represents the concentration of the atacicept and B1y§ complex, plotted against the time, expressed in days, for subcutaneous administration, the main pharmacokinetic measurement. Each line in the graph represents a dose, as presented in the explanations of FIG. one.

FIG. 3A and 3B graphically represent the biological effects of subcutaneous administration on the levels of various immunoglobulins and B-cell levels.

FIG. 4 shows the difference in bioavailability, as shown by measurements of free ataccept for subcutaneous and intravenous administration of ataccept.

FIG. 5 graphically represents the relative similarity of biological activity that was observed using two methods of administration, as represented by a concentration of 1dM versus time. See explanations in FIG. 4.

FIG. 6 graphically shows the similarity of the subcutaneous and intravenous route of administration, as represented by the level of free atacicept versus time.

FIG. 7 shows the relative similarity of the target binding curves that were observed using two methods of administration, as represented by the level of the atacicept-B1y§ complex versus time. See explanations in FIG. 4.

FIG. 8 graphically shows that multiple doses provide higher biological activity, as represented by a concentration of 1dM versus time.

FIG. 9 graphically shows that target binding is higher when using multiple doses, as represented by the level of the ataccept-B1y§ complex versus time. See explanations in FIG. 8.

FIG. 10 graphically represents the concentration of free atacicept, plotted against time expressed in days, for intravenous administration, the main pharmacokinetic measurement. Each line in the graph represents a dose, as presented in the explanations.

FIG. 11 graphically represents the concentration of the atacicept-B1y§ complex, plotted against the time expressed in days, for intravenous administration, the main pharmacokinetic measurement. Each line in the graph represents a dose, as presented in the explanations of FIG. 10.

FIG. 12 is a graphical representation of a biomarker measurement using intravenous administration, in particular, immunoglobulin levels and B cell levels are presented.

FIG. 13A and 13B are graphical representations of the concentration of an atacicept mixture (composite) (defined as a free atacicept + ataccept-B1y§ complex) versus time for subcutaneous administration (study 1). (A) a single dose administration group; (B) multiple dose administration groups. Mean values ± standard error values are presented. Multiple doses were administered on days 0, 7, 14, and 21. The time points for dose administration are not associated with the indicated concentration peaks that were not recorded between doses.

FIG. 14A and 14B are a graphical representation of the concentration of an atacicept composite (defined as a free atacicept + atacicept-B1y§ complex) versus time for intravenous administration (study 2). (A) a single dose administration group; (B) multiple dose administration groups. Mean values ± standard error values are presented. Multiple doses were administered on days 0 and 21.

FIG. 15A-15C are the total profiles of immunoglobulins in Study 1 (subcutaneous injection of 022911), which are demonstrated by the group (% of baseline, mean ± standard error). (A) Σ§Μ; (B) Σ ^ Α, (C) 1dO. These figures extend the data shown in FIG. ZA and ZV.

FIG. 16A-16C are the total immunoglobulin profiles in Study 2 (intravenous administration), which are demonstrated by the group (% of baseline, mean ± standard error). (A) 1dM; (B) 1dA, (C) Σ§0. These figures extend the data shown in FIG. 12.

FIG. 17A-17C graphically represent the relationship between the subcutaneous dose of ataccept (study 1) and the observed maximum immunoglobulin response (in% reduction from baseline). Columns represent mean ± standard error. (A) 1dM; (B) 1dA, (C) 1dC.

FIG. 18A-18C graphically represent the relationship between the intravenous dose of ataccept (study 2) and the observed maximum immunoglobulin response (in% reduction from baseline). Columns represent mean ± standard error. (A) 1dM; (B) 1dA, (C) 1dC.

FIG. 19A and 19B show Σ§Μ profiles (mean ± standard error) in the administration groups of the same single dose of studies with subcutaneous and intravenous administration. (A) 3 mg / kg; (B) 9 mg / kg.

FIG. 20A and 20B show profiles of the atacicept-B1u8 complex (mean ± standard error) in groups of the same single dose of studies with subcutaneous and intravenous administration. (A) 3 mg / kg; (B) 9 mg / kg.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a method for treating a patient suffering from a Σ or ΣΣ stage of systemic lupus erythematosus, which comprises administering weekly a dose of from about 1 to about 10 mg / kg for about 2 to about 52 weeks of a fusion molecule containing a ΤΑCΣ extracellular domain or a fragment that binds to B1u8 and the constant domain of human immunoglobulin.

In a preferred embodiment of the claimed invention, said ΤΑCΣ extracellular domain has a sequence comprising 8EO ΣΌ N0: 1, or said ΤΑCΣ extracellular domain is at least 50% identical to 8E0 ΣΌ N0: 1.

It may also have a sequence comprising only amino acids 30-110 8E0 ΣΌ N0:

one.

The specified constant domain of a human immunoglobulin in a preferred embodiment has a sequence comprising 8E0 ΣΌ N0: 2.

In a most preferred embodiment of the method, said fusion molecule is an ataccept.

A fusion molecule may be administered to a patient in an amount of from about 1 to about 9 mg / kg.

The method in accordance with the invention provides for the additional administration to the patient of a second drug, which is selected from the group consisting of K8AJ, antimalarials, corticosteroids, immunosuppressive agents, ΣνΣ§, ΌΗΕ and thalidomide.

Preferred embodiments of the invention contemplate any suitable route of administration, including subcutaneous, oral or intravenous. In a preferred embodiment, the patient is a human.

B cells are now thought to play an important role in the pathogenesis of 8E using antibody-dependent and antibody-independent mechanisms. In addition to generating antibodies, B cells secrete many cytokines, act as antigen-presenting cells, and perform a variety of effector functions. Thus, B-cells act as rational targets for the development of drugs for SbE (Wto ^ spd 1.B., No. 1 Ksu Aegid Όίδοον 2006; 5: 564-76).

Several B-cell strategies have been proposed as possible therapies for 8E. Some of these strategies are designed to eliminate B cells through the use of monoclonal antibodies (TAB) (Lebbgo Μ.Σ., Eb \ uyB8 1.C., Satbbde Ο.Σ. EbegMet Μ.Κ., Pebet E.A. Aybpek Keit 2002; 46: 2673-3; Lopso Kp., Apoc Σ.Η., Satbc11 Ό.Σ., Re1dag K.E., Woopd R., Atepb L.E., e! A1., Amlb18 Keit 2004; 50: 2580 -9; Leapbgos Μ.Σ., Satbpbde O., Efuagbz 1.S., Ebgep8! E1n M.K., Σδepbegd E.A., Kheita! Od1od 2005; 44: 1542-5; Eotpeg T., KaiGtap 1 ., Uedepeg, U.A., Teoy Ν, OoIepett Ό.Μ., Viggs81sg O.K., Ar1bO8 Ke8Tet 2006, 8: K74). At the same time, others impede the stimulation of B-cells (Vakeg K.R., Eb \ uyB8 Β.Μ., Μa ^ η 8.Η., Cbo1 O.N., Uadeg K.E., Na1regp U.O. , e! a1. Am1bt18 Keit 2003; 48: 3253-65; \ ya11ass Ό.Σ., 8888e Σ., 8! b1 U., ΜcKau Σ., Bbpd E1 Μе ^^^ 11 1.T., e! A1., Atepsap Sodede oG Keita! o1od Appia1 8s1epPpps ΜeeΜ ^ ηд, 2006; Ogo88 1.A., EShop 8.K., Μπάτί 8., 1oЬп8! op Σ., Yyai A., Kossche K., e! a1 ., LptipPu 2001; 15: 289-302) or seek to selectively target B cells producing autoantibodies (Α1а ^ сοη-8едоν ^ а Ό., Титъп 1.A., Риге К.А., ΜсКау Σ.Ό. , SatFe1, Μ.Η., 8ΐππιά V., e! A1., Atlbt18 Keit 2003; 48: 442-54; Beat Ό., Eauap Μ., 21pdet Η., Ni Σ.Ρ., Ζοζе8 Е. 1. Спп Σттиοο1

- 3,029,911

2004; 24: 579-90; Maiegtapp Ν., §1 ЬедедегедΖΖ Ζ., Ζίη§βΓ Η., Μοζβδ Ε., C1t Εχρ 1type1 2004; 137: 513-20).

Attempts to inhibit B-cell stimulation have focused primarily on receptor-ligand interactions that involve molecules called the Wlimphocyte stimulator (B1y§) and the proliferation-stimulating ligand (APME). B1y§ and APP1b are members of the family of tumor necrosis factor (ΤΝΡ) cytokines that are important for the survival of B cells and their development after leaving the bone marrow. B1y§ and APME bind to the same and different receptors. Both molecules bind to the transmembrane activator and partner (TAC1) of the calcium modulator and cyclophilin ligand (CAM) and the Cell Maturation Antigen (BCMA), while B1y§ also binds to the B cell activating factor, which belongs to the receptor of the ΤΝΡ family ( BAPP-M), and APME interacts with proteoglycans.

The evidence obtained in animal models and in humans supports the important role of B1y§ and APME in the development of autoimmune diseases. Transgenic mice that overexpress B1y§ demonstrate B-cell proliferation and polyclonal hyperglobulinemia (Ogo88 TA., 1oNp81op 1., Mibb δ., Epgeletop K., Όί11οη δ., Mabbep K., e! A1., ShOie 2000 : 404: 995-9; Maskau R., Hoobskos 8.A., Bate! Op R., Atbose S., Bae18siet M., 8sbn1bet R., e! A1., 1. Εχρ Meb 1999; 190: 1697- 710; Kyate δ.Ό., 8ato51 I., X1a Χ.Ζ., MsCae δ., Mshet K., 8o1ouuuu I., e! A1., Prgos Νηΐ1 Asab 8sh 2000; 97: 3370-5). Some of these mice develop a lupus-like phenotype consisting of antibodies directed against double-stranded DNA (dsDNA), immunoglobulin deposition in the kidneys and accelerated development of glomerular disease, B1u8 levels are also elevated in lupus-prone ΝΖΕΙΝΖΧν P1 (ΈΙ \ ν) and MKE -1rt / 1rt of mice (δ! Ο11 V., Chi Ό., K1sh K.8., Ko88 M.K., 1dep8e8 Τ.Ν., Yeosyagap V., e! A1., At1yt18 Kueit 2005; 52: 2080- 91). Human studies also suggest the role of B1u8 and APME in systemic autoimmune diseases. Patients with 8LE had elevated serum B1u8 levels, which positively correlated with the levels of anti-dsDNA antibodies (ENAPD 1., P.O8sike V., Vakeg K., νηπβ Ζ., A1agcop δ.δ., Pe881eg B.1. , e! a1., 1. 1ttypo1 2001, 166: 6-10; Sjöeta Ο.δ., R.o8sike V., Nbye I.M., δ! ο11 V., Atlbt18 Iyeit 2001; 44: 1313-19 ; δ! ο11 V., Ме1ua8 δ., Tap 8.M., Sjöta Ο.δ., Oatag V., Chi I., e! a1., Am1b1818 Vkeit 2003; 48: 3475-86). Serum levels of APM are elevated in patients with δΕΕ compared to healthy individuals and in patients with rheumatoid arthritis (Kouata T., T8ikato! O N., M1uad1 Υ., Iter I., O! 8ika 1., Muadata N., e ! a1. App Iyeit Όί8 2005; 64: 1065-7). Serum levels of B1y8 and APM were determined in the synovial fluid of patients with inflammatory arthritis (Tap 8.M., Chi I., Vo8skke V., Repu IV., AgkGe1b Ό.Ο., ^b ^ e8taηη Ο.Κ. e! A1. , Spb18 Vkeit 2003; 48: 982-92). Such convincing observations in mice and humans led to the development of several B1u8 antagonists. One of these agents is a recombinant fusion protein comprising the extracellular domain of the TAC1 receptor linked to the human 1dO1 Pc domain (ataccept, previously designated as TAC1-1d). Ataciceptus blocks the stimulation of the Cells with both B1u8 and APME. Several lines of research provide support for the assumption that atacicept will have powerful effects w νίνο. First of all, transgenic mice that express atacicept have a small number of B cells and reduced concentrations of immunoglobulin, and treating them with atacicept slows the onset and reduces the severity of arthritis in a mouse model of collagen-induced arthritis. The production of antibodies against collagen is also suppressed (Ogo88 TA., IShop δ.Ρ .., Mibb δ., 1oNp81op 1., Ynai A., Vosche M., e! A1., Above). Thirdly, treatment of lupus-prone female ΡΙ \ ν mice with atacicept slows down the development of proteinuria and increases survival (Ogo88 TA., 1oBn8! Op 1., Mibb δ., Εη5е1taη M., Ishop δ., Mabbep K., e! A1., above). In conclusion, by directly comparing the efficacy of mouse atacicept and BAPP-RPD (B1u8 inhibitor only) in lupus-prone female ΕΙ \ ν mice, only atacicept reduces serum levels of 1dM, reduces the incidence of antibody-forming cells in the spleen, and inhibits the 1dM response to antigen, dependent on T-cells, which makes it possible to assume the role of APM in these processes (Watapi) at M., \ ν3ΐ ^ X., Niapd V., Lw Ζ., 8c1iGGsg b., Tao N., e! A1., 1. S’Pp 1n \ 'S81. 2006; 116: 724-34). In the light of these encouraging preclinical data, the inventors tested the biological effects, pharmacokinetics, pharmacodynamics, and safety of ataccept in clinical trials in patients with δ6Ε in accordance with this application.

In both phase Ι studies described in Examples 1 and 2 below, ataccept was well tolerated locally and systemically in patients with δΕΕ. Obvious signs of the biological activity of atacicept were observed with these promising indications, largely in accordance with its mechanism of action (MoA).

In accordance with modern concepts regarding the mechanisms of action (MoA) for ataccept and without linking it with theory, we can say that inhibition of B1u8 and APME leads to effects on B cells, including the secretion of non-specific and specific antibodies, which mainly affects bound with δΕΕ biomarkers and markers of clinical efficacy. As this is typical of phase I studies, attention in this analysis focused on the early stages of the MoA cascade and, in particular, on responses starting with the early biomarkers B1u8 and APME inhibition (such as the atacicept-B1uδ complex) and biological effects (such as as levels 1d).

- 4,029,911

Atacicept demonstrates multiphase nonlinear RK, characterized by a more than proportional dose increase in the effect of the free drug, and an excess increase in the effect of the ataccept-B1y§ complex. This behavior was not unexpected and was shown in CA patients. This supports the hypothesis that ataciccept RK is mediated by its ligands. The general value of ataciceptum RK, although linear, is predictable for a combination of doses, between single and multiple doses. Three PK of atacicepta markers had similar behavior in CA and 8BE patients, which indicates that the type of autoimmune disease is not the main determinant of PK of ataccept.

Prolonged accumulation of the atacicept-B1y§ complex with multiple doses (up to four weekly doses of groups 5 and 6, study 1), together with minimal accumulation of free ataccept, provides confirmation of the presence of a significant initial load of free B1y§ and ARK1b, both systemic and peripheral. Elevated baseline levels of B1y§ measured in three studies (compared with normal subjects, data for which were taken from the literature), speak in favor of this hypothesis.

It is also likely that if the balance between soluble ligands and their receptors existing prior to dose administration is disturbed by administration of an ataccept, complex kinetic processes of redistribution by the blood circulation system, lymphatic system and peripheral compartments are initiated. This redistribution involves both the drug and its ligands, and taking into account the size of the molecule involved, it takes at least several weeks to establish a new equilibrium.

On the other hand, long-term accumulation of the complex may suggest endogenous generation of free ligands (again, both in the blood circulation system and in peripheral tissues). Published data on the rate of increase in serum B1y§ level after administration of retuximab (SatLpbde c1 a1., AgOlPN Kjeit 2006; 54: 723-732) provide additional evidence that the endogenous formation of B1y§ plays an important role in B1y§ inhibition and should be taken into account . A long period of reaching a stable state (within one month of a weekly dose) supports these hypotheses.

Saturated atacicept kinetics were first observed, and were reported using single doses of ataccept, applied to healthy volunteers and CA patients in previous phase I studies, they showed that B1y§ (and ARK1b) inhibition is saturable, i.e. an increase in the effects of ataccept within the saturation point will lead to a decreasing effect in relation to the binding of B1y§ (and ultimately, ARK1b). This phenomenon should be taken into account and used when choosing therapeutic dosage regimens.

It should be emphasized that an acceptable saturation of B1y§ inhibition (and ARK1b) requires maintenance for a certain time and will be achieved using an acceptable model of the effects of atacicept in time. The latter will be necessary for a dynamic balance between the complex and mainly uncharacterized processes of the formation and redistribution of endogenous B1yg and ARK1b and the formed kinetic profile of the atacicept. Such a balance can only be achieved using an acceptable model of the dosing regimen in relation not only to the dose level, but also to the frequency of dosing.

Well-defined relationships between the cumulative dose of atacicept and the 1d antibody response were established using uninsulated placement methods; this relationship was first determined using single doses of atacicept in healthy volunteers and using single and multiple doses of atacicept in patients with CA. In the present studies, all three 1d markers used for monitoring showed a rapid decrease after the first dose of atacicept was administered. After four weeks of dose administration, all three biomarkers of antibody genesis gradually, sequentially and systematically decreased towards a stable state, apparently in the absence of reaching it during the period of dose administration.

The discovery of the fact that the frequency of administration of the dose turns out to be such that it plays at least an important role as the dose level in the response of all three biomarkers, first made in the study of CA, is confirmed by 8E data obtained after subcutaneous administration (study 1) . In general, biomarkers have very similar behavior at similar dose levels in both the 8E and CA populations, which emphasizes the presence of common roots in MoA for both indications, based on the inhibition of B1yA (and ARK1b).

Other interesting facts were revealed from a comparison of PK and the results of biological activity between the two studies. Despite the fact that none of them was intended to address the question of the suitability of the subcutaneous route of administration, to compare the partial and total areas under the concentration-time curves (AIS) of the free atacicept and ataccept compound after administration of such subcutaneous and intravenous doses (group 3, study 1 against group 1 of study 2, and group 4, study 1, the prototype of group 2, study 2) allows us to differentiate a rough estimate of the average bioavailability. From the table. 2ab and 3ab, these estimates are approximately 35–40% for both free and composite drugs — a number that does not go beyond the range of figures for molecules with high molecular weights (Royet and Syattai, 1. Ryatt δοί 2000; 89: 297-310).

However, observation of markers of biological activity revealed that similar doses provide similar biological activity regardless of the route of administration, as illustrated for 1dM in FIG. 19. At first glance, the observation that a 2.5-3-fold difference in the systemic effects of the drug can lead to similar biological effects is contrary to the established system of concepts; however, in the case of ataccept, this phenomenon can be well justified.

A modern understanding of the absorption of large protein molecules after subcutaneous administration indicates that there is an outflow of proteins from the injection site, both in the peripheral lymphatic system and in the blood capillaries, and absorption by the lymphatic system increases with increasing molecular size. For drugs with a molecular weight comparable to that of atacicept, it can be expected that up to 70-80% of the subcutaneous dose can supposedly first enter the peripheral lymphatic system. However, the blood circulation system and the lymphatic system are closely intertwined and connected so that the mass exchange between the blood and lymph compartments will be very quick and unhindered even for large molecules. The latter was confirmed in the case of ataccept by using relatively (for a molecular weight of 73.4 kDa) rapid balancing of PK profiles for intravenous and subcutaneous administration.

These considerations lead to at least two possible, interrelated and, therefore, in no way mutually exclusive explanations of the observed phenomenon. A kinetic explanation suggests that even with subcutaneous administration, a sufficient amount of the drug is transferred to the blood circulation system to ensure adequate inhibition of B1y§ and, thus, to start the MoA cascade in the central compartment. It is well known that many biological effects slow down in relation to the underlying kinetics of the drug. Despite the fact that in the case of ataccept, RE 1ad is not excessive (as evidenced by the rapid decrease in the level of 1d markers after the first dose), this is sufficient to ensure RK 1ad caused by almost inappropriate absorption. This hypothesis is supported by almost identical profiles of the atacicept-B1y§ complex in groups of the same single dose with subcutaneous and intravenous administration (Fig. 20), where similarity, in particular, is noted in the first 7 days after administration.

The pharmacodynamic explanation is that both the blood circulation system and the lymphatic system are sites of action for the inhibition of B1y§ (and ARK1b) and, as such, are targets for atacicept. With the intravenous route of administration, the drug is first injected into the bloodstream and from there it is distributed to the lymph and other (target and non-target) peripheral tissues. In the subcutaneous route of administration, the drug first undergoes outflow into the lymphatic compartment and into the bloodstream in parallel and from the latter is distributed to other (target and non-target) peripheral tissues. In both cases, the penetration of the drug into both sites of action is immediate and rapid and leads to similar profiles of biological activity in each of them.

This interesting fact confirms the hypothesis that the mechanistic application of the knowledge system, which prescribes that the assessment of the effect of protein drugs administered subcutaneously, which should be carried out using the systemic or serum bioavailability parameter, may be unacceptable or, at best, incomplete.

As a rule, the higher the system availability, the greater the effect may have important exceptions in this class of drugs.

The latter has important practical applications related to the development of a therapeutic regimen using atacicept. It becomes clear that the intravenous route can only be a means for delivering higher doses of the drug to the patient, if necessary, provided that the subcutaneous dose can be limited by the volume of injection and the concentration of the solution containing the dose of the drug.

The good tolerability, pronounced biological activity of the treatment with atacicept along with its MoA and other positive patterns observed in the two phase I studies of 8E, provide a reasonable explanation for further research work in relation to the drug for the treatment of patients with 8E. In accordance with the modern system of views on the development of a medicinal product, at each stage, the newly obtained information should be appended to existing information, while the knowledge bank regarding the medicinal product is updated, expanded and improved for subsequent use in order to create a model based on the available information. stages in a typical cycle to study and confirm. In accordance with such a system of concepts, the inventors preferred to observe and analyze many effects (free and composite ataccept), specific binding (ataciceptV1u§ complex), biological activity (1d and the number of cells of the immune system) and certain markers associated with the disease (anti-dsDNA antibodies) in two early studies using

- 6 022911 of a very complex model (sequential, with increasing dose). By analyzing the value of data obtained scientifically and extracting the information that it contains, the authors were given the opportunity to determine the dose intervals and modes for further studies that will be needed to characterize the safety profile of atacicept, and in order to improve understanding of its MoA , confirm the initial clinical efficacy data and determine the optimal clinical application.

Example 1. Subcutaneous administration of atacicept.

This phase 1b, double-blind, placebo-controlled study with an increase in dose included six groups (n = 8 each, except for group 5, in which n = 7) of patients who were treated with atacicept or placebo in a 3: 1 ratio . Groups 1-4 received a single subcutaneous dose of placebo, or 0.3, 1, 3, or 9 mg / kg of atacicept. Groups 5 and 6 received four weekly placebo doses or 1 or 3 mg / kg of atacicept (see Table 1). Patients were followed up for 6 (groups 1-4) or 9 (groups 5 and 6) weeks. Measurement results included: (ί) systemic and local tolerance of ataccept; (i) the incidence of side effects (AE); (ίίί) pharmacokinetics and pharmacodynamics of ataccept, including effects on lymphocyte subpopulations and levels 1§; and (ίν) measuring 8BE disease activity.

Patients with mild to moderate S8E were listed. The biological activity of atacicept was demonstrated by dose-dependent reductions in immunoglobulin levels and the number of mature B cells and the total number of B cells. This effect was most powerful in the repeated dose groups and did not fade over the entire subsequent period. No changes were observed in the number of T cells, natural killer cells or monocytes. Mild reactions at the injection site occurred more frequently in the atacicept group than in the placebo group. No differences were observed in the frequency or type of side effects, and there were no severe or serious side effects in patients who were treated with atacicept.

Pharmacokinetics were evaluated by measuring serum levels of free atacicept (Table 2a), atacicept / B1u8 complex (Table 3a), and composite ataccept (defined as free ataccept + ataccept-B1u8 complex, table 4a). Serum levels of each of the above were quantified using enzyme-linked immunosorbent assay. Serum was incubated with murine TAB conjugated with biotin and specific for ataccept (determination of free or total ataccept) (2utoSepeys8, 1ps., 8caO1s. \ UA). or with goat polyclonal antibodies conjugated with biotin and specific for either B1u8 or ataccept (definition of the ataccept / B1u8 complex) (К & Ό 8у81ет8, М1ппэаро118, ΜΝ) immobilized on a Microplate coated microplate (M1). Antibodies were incubated together with patient samples, standard and control samples diluted 1:10 for 1 h. After washing, the atacicept-specific mouse TAb was conjugated to horseradish peroxidase (NCR) (to measure free atacicept or ataccept-B1u8 complex) , or in the case of the EB18A composite, the anti-ataccept and B1y8 composite (2utoSepaise8, 1 ps) were added and incubated at room temperature for 1 h. In all three analyzes, serum levels of ataccept were determined and quantified using standard chemiluminescence methods, i.e. after washing, tetramethylbenzidine (TMB) was added as NKR substrate (81dta-A1Ftsy, 81. Loshk, MO). The reaction was stopped after 20 minutes using 0.5 M sulfuric acid and the absorbance was recorded at 450 nm.

The analyte concentration in the patient sample was recounted using a standard curve using a second-order alignment algorithm for the polynomial. All samples were measured in triplicate.

Analytical performance criteria accurate to <15% coefficient of variation (CS) for standard samples and <20% for patient samples were acceptable.

The lower limits of quantitative assessment (BBOO) of the analyzes were 15.6 ng / ml for free atacicept, 5 units / ml for the ataccept-V1u8 complex (1 unit / ml corresponds to 1.82 ng / ml of ataccept - 0.44 ng / ml B1u8 in a molar ratio of 3: 1) and 25 ng / ml for composite analytes. The average recovery value of the added analyte, determined for the accuracy analysis for low, medium and high analyte concentrations in patients' CA samples, corresponded to recovery values of 82.5-97.0, 93.9 and 102.0-125.8% in three analyzes, respectively. Serum RK markers were tested as described below (ί) for a single dose of group 1-4 - at a basic level and after 4, 8, 12 hours on the day of administration and after that on study days 2, 3, 4, 8, 15, 22, 29 and 43; (ίί) for a multiple dose of groups 5 and 6 at a basic level and after that on study days 8, 15, 22, 29, 36, 43, 64. In all RK groups, samples on the day of dose administration are indicated at least on the graph.

Unbound B1u8 concentrations were measured in serum at baseline. B1u8 was measured with E18A. Biotinylated TAb specific for B1u8 was incubated with patient samples, standard or control samples (diluted 1:10) for 1 h in microplates pre-coated with streptavidin. After washing with anti-B1y8 NKR, conjugated mouse TAB was incubated at room temperature for 1 h. After washing, TMB was added as an NKR substrate. The reaction was stopped after 20 minutes using 0.5 M sulfuric acid and the absorbance was recorded at 450 nm. The analyte concentration in the patient sample was recounted using a standard curve using a second-order alignment algorithm for the polynomial. All samples were measured in triplicate. Analytical performance criteria accurate to <15% coefficient of variation (CS) for standard samples and <20% for patient samples were acceptable. The lower limit of quantification (LbP) was 15.6 ng / ml B1y3 in serum. The average recovery value of the added analyte for low, medium, and high analyte concentrations in patients' CA samples corresponded to recovery values of 101-113%.

Pharmacodynamics were evaluated by measuring serum levels of immunoglobulins (1dO, 1dM, 1dA), complement-3 (C3) and antinuclear antibodies (ΑΝΑ), and by flow cytometric analysis of lymphocyte subsets. Immunoglobulins and C3 were measured using standard methods. ΑΝΑ were measured using the A1yeya MiShaia1u1e ΑΝΑ analytical system (Ζοιΐ5 8с1еййс 1пс, Katyai, N1, and IL). 1dO, 1dM, and Ι§Α were evaluated in the blood as markers of biological activity. Biomarkers were measured at baseline and after 8 hours on the day of administration (groups 1-4 only) and after that on study days 8, 15, 22, 29, 36, 43 and 64.

The panel of peripheral blood mononuclear cell types (subsets of B and T cells, natural killer cells [ΝΚ] and monocytes) was evaluated in peripheral blood samples stained with antibodies using four-color flow cytometry. The analysis included: summed T cells (SI45 +, SI3 +), T-helper cells (SI45 +, SI3 +, CB4 +, SI8-), Ttitotoksicheskie / suppressor cells (SI45 +, СО3 + .СЭ4-. СЭ8 +). summarized B cells (SI1 9+), mature B cells (SI1 9+, 1dI +, SI27-), monocytes (SI45 +, SI3-, SI14 +, SI56-) and ΝΚ cells (SI45 +, SI3-, SI14-, SE56 +). The organization with which the contract was concluded (EyUypkh, Otoshidei, Thieu No. 1yet1aib§) carried out the processing of samples, staining with antibodies and acquisition, analysis and quality control of data. The authors carried out further analysis and quality control on subsets of B cells. For a subset of B cells, the analytical filter was increased to include small and large lymphocytes, with the latter that were similar in size to monocytes.

Medical records were collected on inclusion on a weekly basis and included in the study. Hematological and blood chemistry profiles were performed weekly and evaluated using the general toxicity criteria of the National Cancer Institute. Blood samples for pharmacokinetic evaluations were collected weekly for repeat dose groups and on day 1 at 4 and 8 hours, as well as on days 2, 3, 4, and 8 and then weekly for single dose groups. Blood samples for pharmacodynamic evaluations were collected weekly for repeat dose groups and on days 2, 3, 8, and then weekly for single dose groups.

An electrocardiogram after day Ό4 was performed once every two weeks in single dose groups and weekly in patients who received repeated doses of the study drug.

Although the study was not aimed at determining the effect of treatment on disease activity, the following measurements were taken to provide preliminary data on efficacy. 3ΕΕΕΝΑ 3ΕΕΌΑΙ scores were determined at the basic level and on days 29 and 43 (groups 1-4), as well as on days 22 and 64 (groups 5 and 6). Antibody and C3 anti-dsDNA levels were measured at baseline, on days 15, 29 and 43 (groups 1-4), as well as on days 15, 22, 29, 43 and 64 (groups 5 and 6).

Methods of data analysis, which included concentration-time profiles, were subjected to non-compartmental analysis (NСΑ; ^ ίηΝοηΣίη software, version 5.0.1). Ignored all measurements below ECOO for NСΑ. Data based on the use of biomarkers (1dM, 1dO or ΙβΑ) turned the format into a difference from the base level and then individual biomarker-time profiles were also subjected to NСΑ. The obtained NC-derived measurements for exposure (RC) and response were sequentially analyzed together to study the existing relationship of response response.

Confirmation of nonlinear pharmacokinetics consistent with the pharmacokinetics of saturable binding of the ligand-receptor interactions was obtained (FIGS. 1 and 2). Multiple-phase pharmacokinetics with clear, fast absorption showed time-concentration profiles for free and composite ataccept, T _max was approximately 24 hours after the first dose, and the initial distribution phase lasted 7-14 days. We observed a low accumulation of free atacicept in repeat dose groups; the accumulation of the composite atacicept was slightly higher, and the ataccept-B1y3 complex was found to undergo accumulation during the dose period.

Treatment with atacicept with an initial, temporary increase in the level of mature and total B cells, followed by a delayed dose-dependent decrease (Fig. 3B). In the single dose groups of 3 mg / kg and 9 mg / kg and in the repeat dose groups, a 35% decrease compared to the baseline in the case of mature B cells was observed on day 29. In the single dose groups, this decrease lasted until day 43; in repeat dose groups, a decrease of approximately 60% was observed per day

- 8 022911

43, and it stabilized by 45-60% until the last assessment on day 64. The models observed for total B cells were similar to those for mature B cells. In the single dose group of 3 mg / kg, a decrease of about 30% from the baseline for total B cells was observed on day 29, and it remained stable until day 43. In the repeated dose groups, a decrease of about 40-50% was observed per day 43, and it stabilized by 35-60% until the last assessment on day 64 (Fig. 3B). There were no significant changes in the number of total, helper or cytotoxic T cells, ΝΚ cells or monocytes.

A dose-dependent decrease in immunoglobulin levels was observed in patients who were treated with atacicept (Figs. 3A and 3B, see also Table 5a). This effect was most significant in repeat dose groups. 1dM levels showed the greatest reduction in treatment, reaching approximately 50% per day 43 in the repeat dose group of 3 mg / kg. 1dA levels decreased by approximately 33% in the repeat dose group of 3 mg / kg on day 29, and 1§C levels decreased by approximately 16% in the repeat dose group of 3 mg / kg on day 36. The lowest levels occurred between days 15 and 29 in single dose groups between days 29 and 43 in repeat dose groups.

After this, the values began to return to the base level. The last observed values were 5-30% below the baseline in the single dose groups (with the exception of the 0.3 mg / kg group in which 1dM values were above the baseline) and 8-65% below the baseline in the repeat dose groups.

These results show that more frequent administration of lower doses of atacicept provides better biological activity than more frequent administration of doses at higher doses (FIGS. 7 and 8).

Example 2. Intravenous administration of ataccept.

This phase 1b, a double-blind, placebo-controlled study with a dose increase included four groups (n = 6 each) of patients who were treated using intravenous atacicept or placebo in a 3: 1 ratio. Groups 1-3 received a single dose of placebo,

3, 9 or 18 mg / kg of atacicept, the second dose was administered three weeks after the initial dose. Measurement results included: (ί) systemic and local tolerance of intravenous atacicept; (i) the incidence of side effects (AE); (ίίί) pharmacokinetics and pharmacodynamics of intravenous ataccept, including effects on lymphocyte subpopulations and levels 1e; and (ίν) measuring the activity of §6E disease. Subjects were evaluated over a period of 6 weeks (groups 1-3) or a period of 9 weeks (group 4); subjects from groups 3 and 4 returned on study days 84 and 120 for sampling on the RK and biomarkers. Serum RK markers were tested as described below (ί) for single dose groups 1 - at the baseline and after 0.25, 0.5 and 4 hours on the day of administration and after that on days 2, 3,

4, 8, 15, 22, 29 and 43; (ίί) for the multiple dose group 4 - at the basic level and after 0.25, 0.5 and 4 hours on the day of the first injection and after that on days 8, 22, 22 (before the second dose and after 0.25 and 0, 5 hours after the second dose was administered), 29, 36, 43, 64. For groups 3 and 4, the measurement of PK was performed on days 85 and 120. In all groups of the Republic of Kazakhstan, samples on the day of the dose were indicated at least on the graph. Unbound B1y§ concentrations were measured in serum at baseline. 1C, 1dM and 1dA were evaluated in blood as markers of biological activity. Biomarkers were measured at baseline and after on study days 2, 3, 4, 8, 15 (groups 1-3 only), 22, 29, 36, 43, and 64 (group 4 only). And in groups 3 and 4, 1d measurements were also taken on study days 85 and 120.

As in Example 1, mild to moderate patients with 8E were listed, and pharmacokinetics were assessed by measuring serum levels of free atacicept (Table 2b), atacicept / B1y complex (table 3b), and ataccept compound (defined as free atacicept + ataccept-B1y§ complex, Table 4b). The biological activity of ataccept was demonstrated by dose-dependent reductions in immunoglobulin levels and the number of mature B cells and the total number of B cells (see Fig. 11, see also Fig. 5b). This effect was the strongest in the repeated dose group and did not fade over the entire subsequent period. No changes were observed in the number of T cells, natural killer cells or monocytes. Mild reactions at the injection site occurred more frequently in the atacicept group than in the placebo group. There were no differences in the frequency or type of side effects, and there were no severe or serious side effects in patients who were treated with atacicept. A comparison between the subcutaneous routes (see Example 1) and the intravenous route revealed very similar pharmacokinetics (non-linear PK mediated by ligands) and a similar PK, which was predictable and consistent with single and multiple doses (Figs. 9 and 10).

Despite the fact that no advantages of intravenous administration were detected (despite the higher bioavailability when using this method of administration, see Fig. 4), these results also support the conclusion that more frequent administration of lower doses of atacicept provides better biological activity than less frequent dosing at higher doses (FIGS. 7 and 8). These results also indicate that despite the lower bioavailability of the drug when using subcutaneous administration, the binding profiles for the two methods are very comparable (see Fig. 7).

- 9 022911

Dosage branches in phase I 8E studies using atacicept Study 1 - subcutaneous administration of atacicept

Table 1

Group Dose Introduction Number of patients I 1x0.3 mg / kg Single dose 6 active doses, 2 placebo 2 1x1 mg / kg Single dose 6 active doses, 2 placebo 3 1x3 mg / kg Single dose b active dose, 2 placebo 4 1x9 mg / kg Single dose 6 active doses, 2 placebo 5 4x1 mg / kg 4 weekly doses (Ο \ ν) 6 active doses, 2 placebo 6 4x3 mg / kg 4 weekly doses (0% ^g ) 6 active doses, 2 placebo

Study 2 - Intravenous Atacicept Administration

Group Dose Introduction Number of patients one 1x3 mg / kg Single dose 5 active doses, 1 placebo 2 1x9 mg / kg Single dose 5 active doses, 1 placebo 3 1x18 mg / kg Single dose 5 active doses, 1 placebo 4 2x9 mg / kg 2 doses with an interval of 3 weeks 5 active doses, 1 placebo

Ο \ ν - every week.

Table 2a

Pharmacokinetic parameters for free ataciceptus obtained from non-compartmental analysis, study 1 Parameters evaluated after the first dose

T. /, (clock) Ttah (clock) Stah (ng / ml) sheet? (mg * h / l) AiSzzb (mg * h / l) 1.0.3 mg / kg group Wed value (thirty) 401 (477) 28 (9.80) 185 (145) 30.4 (29.4) 17.6 (10.6) Averaged. 204 24 136 19.1 14.1 Group 2, 1 mg / kg Wed value (thirty) 452 (219) 18.7 (8.26) 821 (525) 108 (38.4) 69.3 (29.1) Averaged. 433 24 666 120 67.9 Group 3, 3 mg / kg Wed value (thirty) 651 (218) 28 (9.80) 2600 (745) 287 (55.6) 218 (47.2) Averaged. 572 24 2830 293 239 Group 4, 9 mg / kg Avg. (thirty) 642 (218) 32 (12.4) 6190 (3200) 634 (141) 520 (157) Averaged 653 24 5370 608 474

Parameters evaluated after the last dose

5.4x1 mg / kg group

Wed value (80) 690 (230) but BUT. 147 (24.1) 56.9 (9.80) Averaged. 729 BUT. BUT. 149 59.3 Group 6.4x3 mg / kg Wed value (80) 472 (157) BUT. BUT. 209 (33.7) 96.5 (34.5) Averaged. 492 BUT. BUT. 202 85.8

LISzb - the area under the concentration-time curve from time 0 hours to time 336 hours; LiSmr - LIS from time 0 h to infinity; C _max - maximum concentration; 8Ό - standard deviation; T _1/2 - the final half-life; T _max - time to reach maximum concentration. The last dose was administered at time 504 hours. N = 6 8 6 patients per group.

BUT. - not evaluated in accordance with the sampling plan after the last dose.

- 10 022911

Table 2b

Pharmacokinetic parameters for free atacicept obtained from non-compartmental analysis, study 2

Parameters evaluated after the first dose

t _th (hours) Ttah (clock) Stah (ng / ml) lispchg (mH’H / L) Ais ₃₃₆ (mg * h / l) 1.3 mg / kg group Wed value (δϋ) 743 (216) 0,500 (0) 39.7 (5.49) 912 (189) 815 (170) Averaged. 642 0,500 38.6 953 848 Group 2, 9 mg / kg Wed value (8C> 722 (146) 0.350 (0.137) 198 (248) 2040 (613) 1930 (565) Averaged. 765 0.250 91.7 1770 1680 Group 3, 18 mg / kg Wed value (thirty) 796 (188) 0.400 (0.137) 289 (91.2) 5010 (743) 4840 (713) Averaged. 702 0,500 257 4880 4730

Group 4.2x9 mg / kg with interval. 3 weeks

Wed value (8ϋ) Averaged. BUT. BUT. 1.15 (1.60) 0,500 140 (23.7) 148 BUT. BUT. 2750 (220) 2620 Parameters evaluated after the last dose Group 4, 2x9 mg / kg interval. 3 weeks Wed value (8ϋ) 748 (92.2) Averaged. 710 504.25 (0) 504.25 109 (22.6) 119 4320 (1020) 4700 4110 (971) 4520

LiSze - the area under the concentration-time curve from time 0 hours to time 336 hours; LYSUT - LIS from time 0 h to infinity; C _max - maximum concentration; 8Ό - standard deviation; T _{1/2 the} final half-life; T _max time to reach maximum concentration. The last dose was administered at time 504 hours. N = 6 8 6 patients per group.

BUT. - not evaluated in accordance with the sampling plan after the first dose.

from non-compartmental analysis, study 1

Table 3 a

Pharmacokinetic parameters for the atacicept composite obtained

Parameters evaluated after the first dose You TSCH Stah AiStg Ais ₃₃₆ (clock) (clock) (ng / ml) (mg "h / l) (mg / l) Group 1, O, 3 mg / kg Wed value (thirty) 3710 (5450) 30 (14.7) 436 (300) 1360 (2120) 75.6 (36.5) Averaged. Group 2, 1 mg / kg 1270 24 324 319 61.0 Wed value (thirty) 807 (515) 16 (8.80) 1160 (597) 610 (370) 168 (47.1) Averaged. Group 3, 3 mg / kg 543 sixteen 963 441 169 Wed value (80) 3040 (2770) 32 (12.4) 3140 (935) 2470 (1620) 400 (69.3) Averaged. Group 4, 9 mg / kg 2580 24 3130 2210 411 Wed value (8ϋ) 878 (256) 32 (12.4) 8890 (4860) 1770 (378) 865 (199) Averaged. 795 24 7150 1780 807

__ Parameters evaluated after the last dose

Group 5, 4x1 mg / kg

Wed value (3ϋ) 1410 (572) BUT. BUT. 1570 (834) 237 (50.4) Averaged. 1610 BUT. BUT. 1490 221 Group 6, 4x3 mg / kg Wed value (thirty) 1500 (545) BUT. BUT. 2530 (1340) 386 (83.4) Averaged. 1520 BUT. BUT. 2070 389

LIS ₃₃₆ - the area under the concentration-time curve from time 0 h to time 336 h; LYSUT - LIS from time 0 h to infinity; C _max - maximum concentration; 8Ό - standard deviation; T _1/2 - the final half-life; T _max - time to reach maximum concentration. The last dose was administered at time 504 hours. N = 6 8 6 patients per group.

BUT. - not evaluated in accordance with the sampling plan after the last dose.

- 11 022911

Pharmacokinetic parameters for an atacicept composite obtained from non-compartmental analysis, study 2

Table 3b

Parameters evaluated after the first dose

T _and (hours) t ¹ shah (hours) (ng / ml) Stork (mg * h / l) AiSzzb (MG’h / l) Group 1, 3 mg / kg Wed value (8ϋ) 2550 (1220) 0.350 (0,137) 57.3 (12.0) 2870 (531) 1180 (184) Averaged. 2050 0.250 55.9 2650 1200 Group 2, 9 mg / kg Wed value (δϋ) 1560 (842) 0.350 (0.137) 348 (362) 4590 (520) 3210 (650) Averaged. 1600 0.250 201 4430 2880 Group 3, 18 mg / kg Wed value (fifty) 1080 (147) 0.250 (0) 411 (77.3) 7470 (1380) 5990 (863) Averaged. 1100 0.250 446 7710 6240

Parameters evaluated after the first dose t _th (hours) Ttah (clock) Stah (ng / ml) AiS, Mr (mg * h / l) AiSzzb (mg’h / l) G group 4, 2x9 mg / kg with an interval. 3 weeks Wed value (80) BUT. 1.05 (1.65) 361 (334) BUT. 4720 (2400) Averaged. BUT. 0.250 251 BUT. 4320

Parameters evaluated after the last dose

Group 4, 2x9 mg / kg with an interval of 3 weeks.

Wed value (thirty) 1480 (750) 504.25 (0) 240 (69.4) 8490 (2010) 6390 (1200) Averaged. 1140 504.25 272 8130 6390

АиС ₃₃₆ - the area under the concentration-time curve from the time moment 0 h to the time moment 336 h; AIS ^ - AIS from time 0 h to infinity; C _{max -} maximum concentration; 8Ό - standard deviation; T _1/2 - the final half-life; T _max - time to reach maximum concentration. The last dose was administered at time 504 hours. N = 6 8 6 patients per group. BUT. - not evaluated in accordance with the sampling plan after the first dose.

Table 4a

Pharmacokinetic parameters for the B1y§-atacicept complex, obtained from non-compartmental analysis, study 1

Parameters evaluated after the first dose * P _g Ttah Stah AiS [nr. AVCzy (clock) (clock) [units / ml] (units) h / l) (units) h / l) Group 1, 0, Zmg / kg Wed value (80) 1240 (656) 364 (126) 161 (91.9) 382 (390) 38.5 (17.8) Averaged. G group 2, 1 mg / kg 1230 336 134 259 33.7 Wed value (δϋ) 821 (704) 260 (124) 269 (89.9) 362 (196) 68.7 (24.1) Averaged. Group 3, 3 mg / kg 512 336 286 319 73.0 Wed value (δϋ) 1250 (517) 728 (330) 316 (30.4) 727 (323) 59.5 (13.3) Averaged. 4.9 mg / kg group 1420 840 315 735 55.9 Wed value (80) 6960 (6550) 700 (223) 369 (66.2) 3680 (2890) 69.9 (16.7) Averaged. 6220 672 388 4330 72.7

Parameters evaluated after the last dose

Group 5, 4x1 mg / kg Wed value (δϋ) 2510 (2330) 756 (92.0) 460 (122) 1700 (1330) 139 (38.1) Averaged. 1970 756 443 1260 134 Group 6.4x3 mg / kg Wed value (80) 7680 (13200) 840 (184) 668 (159) 8050 (13700) 201 (40.6) Averaged. 2220 840 660 2450 199

AIS ₃₃₆ - the area under the concentration-time curve from time 0 hours to time 336 hours; AISut - AIS from time 0 h to infinity; C _max maximum concentration; 8Ό - standard deviation; T _1/2 - the final half-life; T _max - time to reach maximum concentration. The last dose was administered at time 504 hours. N = 6 8 6 patients per group.

* T _1/2 and AISut are not reliable estimates for this value due to the terminal shape of the profiles.

- 12,029,911

Pharmacokinetic parameters for the B1y§-atacicept complex, obtained from non-compartmental analysis, study 2

Table 4b

Parameters evaluated after the first dose * t _th Too Wow AiSzzb (clock) (hours) [ked./ml] (ked. "h / l) (units “h / l)

Group 1, 3 mg / kg Wed value (80) 86,500 (159,000) 672 (206) 0.297 (0.0412) 35100 59.6 (10.2) (63800) Averaged. Group 2, 9 mg / kg 9800 672 0,300 4240 63.1 Wed mean, (8ϋ) 3790 (2320) 605 (255) 0.352 (0.0763) 2040 (1230) 61.0 (11.4) Averaged. Group 3, 18 3770 504 0.342 2480 58.8 mg / kg Wed value (8E) 1800 (616) 672 (206) 0.454 (0.118) 1460 (674) 86.3 (20.0) Averaged. 1570 672 0.452 1210 84.3

Group 4.2x9 mg / kg with interval. 3 weeks

Wed value (3ϋ) BUT. 504 (0) 0.414 (0.121) BUT. 78.8 (17.8) Averaged. BUT. 504 0.397 BUT. 78,4 Parameters evaluated after the last dose Group 4.2x9 mg / kg with interval. 3 weeks Wed value (5ϋ) 2650 (3270) 1008 (0) 0.707 (0.343) 2790 (2550) 186 (38.0) Averaged. 1140 1008 0.685 1740 195

LISzb - the area under the concentration-time curve from time 0 hours to time 336 hours; LYSUT - LIS from time 0 h to infinity; C _max - maximum concentration; 8Ό - standard deviation; T _1/2 - the final half-life; T _max - time to reach maximum concentration. The last dose was administered at time 504

h. N = 6 §ЬЕ patients per group.

BUT. - did not evaluate * T _1/2 and LYSut are not reliable estimates for this value due to the terminal shape of the profiles.

- 13,029,911

Table 5 a

Non-compartmental analysis results for (1e) M, 1§L, and 1§O biomarkers study 1

ΙίξΜ 1eA ΙβΟ Group * TPSHL (days) Max, Ttah Max. Ttsuh Max. cheating (% (days) (% of bases. (days) (% of bases. from the bases. level) level) level) Group 1.0.3 mg / kg subcutaneous. Avg. 14.1 (17) 2.82 (23.4) before. Before. 11.8 (15.7) -0.377 (17.8) (80) Averaged. 7 10.2 Group 2, 1 mg / kg subcutaneous. before. before. 7.00 4.07 Avg. 18.7 (13.0) 19.4 (8.93) before. before. 25.7 (14.5) 11.1 (5.77) (51 ” Averaged. 17.5 16.5 Group 3, 3 mg / kg subcutaneous. before. before. 24.5 12.1 Avg. 23.3 (5.70) 24.8 (3.81) Before. before. 29.2 (8.20) 10.5 (1.25) (8E) Averaged. 21.0 25.2 Group 4, 9 mg / kg subcutaneous. before. before. 28.0 10.9 Wed value 29.2 (8.20) 37.9 (8.57) Before. before. 18.7 (7.20) 14.7 (5.95) (5Ώ) Averaged. 28.0 37.2 Group 5, 4x1 mg / kg subcutaneous ¢)% ⁷ BEFORE. before. 21.0 15.9 Avg. 31.0 (11.3) 29.2 (10.0) 27.0 23.1 (10.1) 25.0 (11.3) 11.5 (5.84) (80) (10.2) Averaged. 35.0 31.0 Group 6 4x3 mg / kg subcutaneous. 28.0 20.7 28.0 14,4 Wed value 42.0 (11.7) 50.4 (7.25) 36,2 34.9 (5.85) 33.8 (6.90) 17.5 (4.22) (80) (2.90) Averaged. 42.0 49.8 Placebo) 35.0 32.9 35.0 17.8 Avg. 13.5 (12.4) 13.7 (15.3) 14.0 9.14 (7.06) 11.0 (11.3) 11.2 (13.5) (80) (9.90) Averaged. 7.00 10.9 10.5 10.6 14.0 9.84

* Patients with an active dose for groups 1-6 (n = 6).

Ψ Placebo group, all placebo patients combined together (n = 12). T _tax - time of maximum exhaustion 1d

before. - no data.

Table 5b

Non-compartmental analysis results for (1e) M, 1§L, and 1§O biomarkers study 2

Group * T _ha „(days) Max. T _max Max. T _max Max.

cheating {% (days) change (% (days) (% of base. of base. of base. level) _level) _level) _

Group 1.3 mg / kg intravenous.

Wed value (3ϋ) 23.8 (12.7) 34.2 (11.2) 18.2 (14.5) 19.0 (10.9) 15.4 (7.70) 13.9 (7.72) Averaged. 21.0 39,4 14.0 14.9 21.0 12,2 Group 2, 9 mg / kg intravenous. Wed value 25.2 (3.80) 33.5 (5.64) 21.0 (7.00) 26.5 (3.43) 21.0 (7.00) 17.1 (3.86) (8ϋ) Averaged. 28.0 33.6 21.0 25.8 21.0 17.1

Group 3, 18 mg / kg intravenous.

Wed value (8E) 29.4 (7.70) 35.3 (13.1) 23.8 (6.30) 29.8 (5.71) 19.6 (7.70) 17.1 (1.96) Averaged. 28.0 29.3 28.0 28.6 14.0 17.0 Group 4.2x9 mg / kg intravenous, with an interval. 3 weeks Avg. 39.2 (6.30) 42.3 (10.7) 36.4 (7.70) 35.9 (8.06) 36.4 (7.70) 23.2 (3.76) (thirty) Averaged. 42.0 41.2 42.0 37.5 42.0 24.0 Placebo! Wed value 10.2 (12.0) 8.51 (5.14) 21.5 (17.3) 15.4 (16.1) 10.0 (12.2) 10.7 (7.14) (5E) Averaged. 5.00 7.66 21.0 10,2 5.00 13.9

* Active dose patients for groups 1-4 (n = 5).

ί Placebo group, all placebo patients combined together (n = 4). T _tax - time of maximum exhaustion 1d

- 14,029,911

References

The references cited in this invention, including patents, published applications, and other publications, are incorporated herein by reference.

Sequence List <110> ZIMODENENIK. INC.

Ares Trading S. A.

<120> METHODS FOR DOSAGE ADMINISTRATION FOR TREATMENT OF AUTOIMMINAL DISEASES WHEN USING A TASTE FILLED PROTEIN, SUCH AS AN ATACYCEPT <130> 07-LDC <150> 60 / 943,618 <151> 2007-06-13 <150> 61/02431 2008 / 1331,031 -01-28 <160> 2 <170> RaGepIP version 3.4 <210> 1 <211> 154 <212> Protein

Me (8eg O1u Bei O1u Arg 8eg Arg Ag Ar £ O1u O1u Ag§ 8eg Arg Wa1 Azr 15 10 15

O1n O1i and O1i and Arg Р е го го О O1n 1 п еи Т р р Ь г г О 1 1 1 У 1 1 А!!! Ag§

25 30

8eg Suz Rgo O1i O1i O1n Tug Tgr Azr Rgo Bei Bei O1u Tg Suz Me!

40 45

5eg Suz Buz Thr Not Suz Azp Ηίδ O1n 8eg O1n Arg Thb Suz A1a A1a 50 55 60

Rye Suz Arg 8g Bei 8g Suz Ag & Buz O1i O1n O1u Buz Rye Tug Azr 65 70 75 80

- 15,029,911

Ηϊ® еи еи Α?? Avr Su® Ne 8eg Su® A1a Zeg Ne Su® C1u Οίη Ηί®

90 95

Prgo Luz O1n Cy® A1a Tug Rye Cyz O1 and A®n Ll® Lye Ατβ Zeg Prgo Ya1 100 105 PO

Azp еи Р го го го го О О и Те и £ Аг £ § § § § § Α>> З

Azp Zeg Azr Azp Zeg O1y Arg Tug O1n O1u Ley O! And Ηί® Ag§ C1u Zeg 130 135 140

C1 and A1a Zeg Rgo A1a Ley Rgo O1u Ley Ly®

145 150 <210> 2 <211> 348 <212> PKT <213> Constant domain of the human immunoglobulin PC5 <400> 2

Me1 A®p A1a Me1 Ly® Aga O1u Lye Suz Su® Wa1 Lye Lye Ley Suz O1u 15 10 15

A1a V®! Pye Ua1 Zeg Ley Zeg O1n O1 and Pe Ηί® A1a O1i Ley Arg Ag

25 30

Rye Arg Age Ala Me1 Arg Zeg Su® Rgo O1i O1i O1n Tug Tgr Azr Rgo 35 40 45

Ley Ley C1y Thz Suz Me1 Zeg Suz Ly® Thg Ne Su® Azp Ηί® C1p Zeg 50 55 60

O1n Agz Th2 Suz A1a A1a Pbu Suz Arg Zeg Ley Zeg Suz Argby® O1u

70 75 80

O1n O1y LU® Rye Tug Azr Ηΐ® Lye Ley Arg Azr Su® Pe Zeg Suz A1a

90 95

Zeg Ne Su® C1u Οίη Ηί® Prgo Luz S1n Suz A1a Tug Rye Su® <31 and Azp 100 105 PO

Bу® bе Аг § ег ег и го уз ег ег ег З З З З З ® ® ® ® ® ®

- 16,029,111

115 120 125

Рgo Suz Рgo А1а Рго С1и А1а О1и О1у А1а Рго Зег Уа1 Рее Рее Рее 130 135 140

Rgo Rgo Lgo Rgo Luz Azr Thr Ley Me1 Not Zeg Αίβ Thr Rgo O1 and Wa1

145 150 155 160

Thz Suz Wa1 Wa1 Wa1 Azr Wa! Zeg ΗΪ3 O1 and Azr Rgo O1 and Va1 b3 Pb

165 170 175

Azp Tgr Tug Va1 Azr C1u Va1 O1 and Va1 From Azp A1a bz Tg bz Rgo 180 185 190

Arg O1 and Cl and O1n Tug Azp Zeg Thb Thug Arg§ Ya1 Ya1 Zeg Ya1 Ley Ty 195 195 200 205

Ya1 Ley Shz O1n Azr Tgr Ley Azp C1y Luz O1 and Tug Luz Suz Luz Ya1 210 215 220

Zeg Azp Luz A1a Ley Rgo Zeg Zeg Zeg Not O1 and Luz Thz Not Zeg Luz A1a

225 230 235 240

Luz O1y O1n Rgo Lg§ O1 and Rgo O1n Wa1

245 250 255

Azr O1 and Ley Tyr Luz Azp O1n Wa1 Zeg Ley Thz Suz Ley Ya1 Luz O1u 260 265 270

Rie Tug Rgo Zeg Azr Not A1a Ya1 <31 and Tgr O1i Zeg Azp O1u O1p Rgo 275 280 285

O1i Azp Azp Tug Luz Tg Tg Rgo Rgo Va1 Ley Azr Zeg Azr O1u Zeg 290 295 300

Pie Pie Lye Thug Zeg Luz Ley Thy Wa1 Azr Luz Zeg Arg Tgr O1n O1n

305 310 315 320

О1у Азп Уа1 Рее Zeg Suz Zeg Уа1 Ме1 Шз О1и A1а Ле ΗΪ8 Азп Шз

325 330 335

Tug Tg O1n bz Zeg bg Zeg bg Zeg Rgo C1b bb

340 345

Claims (11)

CLAIM 1. A method of treating a patient suffering from stage I or II of systemic lupus erythematosus, including weekly administration at a dose of from about 1 to about 10 mg / kg for about 2 to about 52 weeks of a fused molecule, which includes: (ί) TAC1 extracellular domain or a fragment thereof that binds to B1y§; (i) a constant domain of human immunoglobulin. 2. The method according to claim 1, where the specified TAC1 extracellular domain has a sequence comprising L T) GO N0: 1. 3. The method according to claim 1, where the specified TAC1 extracellular domain is at least 50% identical 5> EO GO N0: 1. 4. The method according to claim 1, where the specified TAC1 extracellular domain has a sequence comprising amino acids 30-110 5> EO GO N0: 1. 5. The method according to claim 1, where the specified constant domain of a human immunoglobulin has a sequence comprising 5> EO GO N0: 2. 6. The method according to claim 1, where the specified fused molecule is an atacicept. 7. The method according to claim 1, where the specified composition is administered in an amount of from about 1 to about 9 mg / kg 8. The method according to claim 1, where this method provides for the additional administration to the patient of a second drug, which is selected from the group consisting of MAY, antimalarial drugs, corticosteroids, immunosuppressive agents, 1U1d, ENEL and thalidomide. 9. The method of claim 8, wherein said fusion molecule is an atacicept. 10. The method according to claim 1, where the specified fused molecule is administered subcutaneously, orally or intravenously. 11. The method according to claim 1, where the patient is a human.